To develop a versatile nuclear-targeted gene vector, nuclear localization signal (NLS) oligopeptides combining cysteine (C), histidine (H), and stearic acid (STR) were investigated in this study. The original SV40 sequence ( SV40 : Pro-Lys-Lys-Lys-Arg-Lys-Val) was selected as the NLS sequence, and physical characterizations of various NLS-based oligopeptides (C SV40 C, STR-C SV40 C, and STR-CH₂ SV40 H₂C), including mean diameter, zeta-potential, complex condensation, and decondensation, were evaluated. In addition, cellular and nuclear uptake of plasmid DNA (pDNA) and gene expression in COS7 and dendritic cells (JAWS II) were determined. As a result, C and STR enhanced formation of a smaller and more stable nano-complex with pDNA based on ionic interactions, the disulfide linkage and hydrophobic interactions. STR-C SV40 C and STR-CH₂ SV40 H₂C had significantly higher cellular uptake ability and transfection efficiency than SV40 and C SV40 C. In particular, STR-CH₂ SV40 H₂C had higher nuclear uptake and gene expression efficiency than STR-C SV40 C. Furthermore, STR-CH₂ SV40 H₂C could deliver pDNA to the nuclei and had high gene expression efficiency in dendritic cells. Our results indicate that STR-CH₂ SV40 H₂C is a promising gene delivery system in non- or slow-dividing cells.